超高效液相色谱-三重四级杆串联质谱法测定贝类中的9种脂溶性贝类毒素
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摘要
目的建立多种贝类食品中9种脂溶性贝类毒素超高效液相色谱-三重四级杆串联质谱法。方法经甲醇提取,以Waters BEH C18(2. 1 mm×10 mm,1. 7μm)为分离柱,乙腈和0. 01%氨水溶液为流动相梯度洗脱,超高效液相色谱-串联四级杆质谱仪测定,外标法定量,测得游离态毒素;采用碱水解将酯化毒素转化为游离态,以Waters BEH C18(2. 1 mm×10 mm,1. 7μm)为分离柱,乙腈和0. 01%氨水溶液为流动相梯度洗脱,超高效液相色谱-串联四级杆质谱仪测定,外标法定量,测得毒素总量。结果本法在2. 5 ng/ml～50 ng/ml内呈良好线性关系,相关系数> 0. 99,最低检出限为10μg/kg～20μg/kg,相对标准偏差<15%。结论本方法操作简单、灵敏度高、重现性好,且具有较好的选择性和特异性,能满足贝类食品中9种脂溶性贝类毒素同时检测的要求,为贝类食物中毒诊断提供了可靠的实验室依据。
Objective To establish a method for 9 kinds of liposoluble shellfish toxins in shellfish by ultra high performance liquid chromatography-triple quadruple mass spectrometry( UPLC-MS/MS). Methods After methanol extraction,the free toxin was separated with a Waters BEH C18 column( 2. 1 mm × 10 mm,1. 7 μm) and detected with triple quadruple mass spectrometer and quantified by the external standard method; acetonitrile and 0. 01% ammonia solution were used as mobile phases. The analytes were detected with triple quadruple mass spectrometer and quantified by the external standard method. The total amount of toxin was measured. Results The method showed a good linear relationship within 2. 5 ng/ml-50 ng/ml with the correlation coefficient greater than 0. 99. The detection limits were within 10 μg/kg-20 μg/kg,with relative deviations less than 15%. Conclusion The method is simple,sensitive,and selective. It can be used for the detection of 9 kinds of liposoluble shellfish toxins in shellfish and provide reliable basis for laboratory.
Objective To establish a method for 9 kinds of liposoluble shellfish toxins in shellfish by ultra high performance liquid chromatography-triple quadruple mass spectrometry( UPLC-MS/MS). Methods After methanol extraction,the free toxin was separated with a Waters BEH C18 column( 2. 1 mm × 10 mm,1. 7 μm) and detected with triple quadruple mass spectrometer and quantified by the external standard method; acetonitrile and 0. 01% ammonia solution were used as mobile phases. The analytes were detected with triple quadruple mass spectrometer and quantified by the external standard method. The total amount of toxin was measured. Results The method showed a good linear relationship within 2. 5 ng/ml-50 ng/ml with the correlation coefficient greater than 0. 99. The detection limits were within 10 μg/kg-20 μg/kg,with relative deviations less than 15%. Conclusion The method is simple,sensitive,and selective. It can be used for the detection of 9 kinds of liposoluble shellfish toxins in shellfish and provide reliable basis for laboratory.
引文
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[13] Morono A,Arevalo F,Fernandez ML,et al. Accumulation and transformation of DSP toxins in mussels mytilus galloprovincialis during a toxic episode caused by Dinophysis acuminata[J]. Aquat Toxicol,2003,62(4):269-280.
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[15]黄聪,李晓晶,彭荣飞,等.固相萃取-高效液相色谱-串联质谱法检测贝类产品7种脂溶性贝类毒素[J].中国卫生检验杂志,2011,21(5):1075-1077.
[16]陈剑刚,朱炳辉,梁素丹,等.固相萃取-液相色谱-串联质谱法测定贝类中的脂溶性贝类毒素[J].中国食品卫生杂志,2015,27(6):624-629.
[17]姚建华.贝类毒素液相色谱—串联质谱检测技术的建立与应用[D].上海:上海海洋大学,2010.
[18]马荣桧,高彦,万进,等.贝类毒素及检测技术的研究现状[J].食品研究与开发,2013,34(22):104-108.
[2] Li A,Ma J,Cao J,et al. Analysis of paralytic shellfish toxins and their metabolites in shellfish from the North Yellow Sea of China[J]. Food Addit Contam Part A Chem Anal Control Expo Risk Assess,2012,29(9):1455-1464.
[3]崔晗,刘玥婷,沈葆真,等.贝类产品中脂溶性贝类毒素检测方法的研究进展[J].食品安全导刊,2017(36):146-147.
[4]谭志军,吴海燕,郭萌萌,等.脂溶性贝类毒素安全评价与检测技术研究进展[J].中国水产科学,2013,20(2):467-479.
[5]刘晓玉.小鼠法检测贝类毒素假阳性结果的来源分析研究[D].大连:大连工业大学,2016.
[6]刘晓玉,徐静,晚观生,等.小鼠法检测脂溶性贝类毒素假阳性来源分析[J].食品科学,2016,37(20):157-161.
[7]赵前程,来鹏,秦成,等. ELISA与小白鼠生物法检测腹泻性贝类毒素的研究[J].食品科技,2009,34(11):287-292.
[8]李军涛,候水平,姬泽薇,等.腹泻性贝类毒素的免疫荧光层析检测方法的初步建立[J].热带医学杂志,2015,15(2):189-192.
[9]方佳如.海洋生物毒素检测的细胞传感器及手持式检测仪的研究[D].杭州:浙江大学,2017.
[10]方丽媛,李代宗,肖勤.贝类毒素及其检测方法研究进展[J].中国渔业质量与标准,2017,7(1):41-49.
[11]李芳,李雪梅,李献刚,等.贝类毒素检测方法研究概况[J].食品研究与开发,2015,36(23):184-186.
[12]刘晓玉,徐静,黄莲芝,等.腹泻性贝类毒素及检测技术研究进展[J].食品安全质量检测学报,2015,6(10):4096-4102.
[13] Morono A,Arevalo F,Fernandez ML,et al. Accumulation and transformation of DSP toxins in mussels mytilus galloprovincialis during a toxic episode caused by Dinophysis acuminata[J]. Aquat Toxicol,2003,62(4):269-280.
[14] Gato-Martinez A,Pineiro N,Aguete EC,et al. Further improvement in the application of high-performance liquid chromatography to the analysis of algal toxins in the aquatic environment[J]. J Chromatogr A,2003,992(1):159-168.
[15]黄聪,李晓晶,彭荣飞,等.固相萃取-高效液相色谱-串联质谱法检测贝类产品7种脂溶性贝类毒素[J].中国卫生检验杂志,2011,21(5):1075-1077.
[16]陈剑刚,朱炳辉,梁素丹,等.固相萃取-液相色谱-串联质谱法测定贝类中的脂溶性贝类毒素[J].中国食品卫生杂志,2015,27(6):624-629.
[17]姚建华.贝类毒素液相色谱—串联质谱检测技术的建立与应用[D].上海:上海海洋大学,2010.
[18]马荣桧,高彦,万进,等.贝类毒素及检测技术的研究现状[J].食品研究与开发,2013,34(22):104-108.